Ostuni Angela, Frontoso Raffaele, Crudele Maria Antonietta, Barca Lorella, Amati Mario, Boni Raffaele, De Vendel Jolanda, Raimondi Paolo, Bavoso Alfonso
Department of Basic and Applied Sciences, University of Basilicata, Via dell' Ateneo Lucano 10, 85100 Potenza, Italy.
Istituto Zooprofilattico Sperimentale del Mezzogiorno, Via Salute, 2, 80055 Portici, Italy.
Pathogens. 2025 Jun 8;14(6):575. doi: 10.3390/pathogens14060575.
Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, often sourced from the Wyoming reference strain. To broaden the range of viral proteins and strains employed in these immunoassays, we previously developed a novel p26/double-strain gp45 indirect ELISA. In this study, we evaluated the performance of this ELISA in comparison to two commercial EIAV ELISAs using Cohen's Kappa test and Bayesian Latent Class Analysis (BLCA), a statistical method that estimates test performance without requiring a perfect reference standard. A comparison with the official classification of the sera by the Italian Veterinary Service was also performed. A total of 372 serum samples, including 96 that were positives by all three tests, were analyzed. Results from both Cohen's Kappa test and BLCA, alongside comparison with official classifications, affirm the diagnostic reliability of the two commercial ELISAs and suggest that the novel ELISA, with its enhanced antigenic diversity, could offer an accurate and reliable diagnostic option for EIAV. This novel assay enhances existing commercial ELISAs and has the potential to strengthen routine diagnostic workflows.
马传染性贫血病毒（EIAV）是一种具有显著遗传变异性的慢病毒，带来了重大的诊断挑战。现有的诊断工具包括琼脂凝胶免疫扩散试验（AGID）、酶联免疫吸附测定（ELISA）和蛋白质印迹法（WB）。ELISA和AGID主要利用p26衣壳蛋白，该蛋白通常来源于怀俄明参考毒株。为了扩大这些免疫测定中使用的病毒蛋白和毒株的范围，我们之前开发了一种新型的p26/双毒株gp45间接ELISA。在本研究中，我们使用科恩卡方检验和贝叶斯潜在类别分析（BLCA，一种无需完美参考标准即可估计检测性能的统计方法），评估了这种ELISA与两种商用EIAV ELISA相比的性能。还与意大利兽医服务局对血清的官方分类进行了比较。共分析了372份血清样本，其中96份在所有三项检测中均呈阳性。科恩卡方检验和BLCA的结果以及与官方分类的比较，均证实了两种商用ELISA的诊断可靠性，并表明这种新型ELISA因其增强的抗原多样性，可为EIAV提供准确可靠的诊断选择。这种新型检测方法改进了现有的商用ELISA，有潜力加强常规诊断工作流程。
